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J Clin Microbiol. Mar 1995; 33(3): 701–705.
PMCID: PMC228017

Development of PCR for screening of enteroaggregative Escherichia coli.

Abstract

In this study, we determined the sequence of the EcoRI-PstI fragment of the plasmid pCVD432, also termed the enteroaggregative Escherichia coli (EAggEC) probe. A primer pair complementary to this probe was designed for PCR amplification of a 630-bp region. Comparison of the analysis of the EAggEC probe sequence with those in database libraries revealed no significant similarity to any known bacterial gene. Pure cultures of E. coli cells, as well as mixed cultures from stool specimens, were investigated with the PCR assay, the EAggEC probe test, and the adherence test. Of 50 E. coli strains which demonstrated aggregative adherence to HEp-2 cells, 43 (86%) were positive with the EAggEC PCR. All 43 of these strains reacted with the EAggEC probe. Six EAggEC strains gave negative results by both molecular techniques. In contrast, only 4 of 418 (0.96%) strains representing other categories of diarrheagenic E. coli demonstrated a positive PCR result. The PCR was also successful in screening for the presence of EAggEC in enriched cultures grown from stool specimens. Compared with cell culture assays and colony hybridization, our findings revealed that the PCR assay was more rapid, simple, and highly sensitive and can therefore be recommended as a screening method for EAggEC in the clinical laboratory.

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Selected References

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  • Albert MJ, Qadri F, Haque A, Bhuiyan NA. Bacterial clump formation at the surface of liquid culture as a rapid test for identification of enteroaggregative Escherichia coli. J Clin Microbiol. 1993 May;31(5):1397–1399. [PMC free article] [PubMed]
  • Baudry B, Savarino SJ, Vial P, Kaper JB, Levine MM. A sensitive and specific DNA probe to identify enteroaggregative Escherichia coli, a recently discovered diarrheal pathogen. J Infect Dis. 1990 Jun;161(6):1249–1251. [PubMed]
  • Bhan MK, Raj P, Levine MM, Kaper JB, Bhandari N, Srivastava R, Kumar R, Sazawal S. Enteroaggregative Escherichia coli associated with persistent diarrhea in a cohort of rural children in India. J Infect Dis. 1989 Jun;159(6):1061–1064. [PubMed]
  • Brian MJ, Frosolono M, Murray BE, Miranda A, Lopez EL, Gomez HF, Cleary TG. Polymerase chain reaction for diagnosis of enterohemorrhagic Escherichia coli infection and hemolytic-uremic syndrome. J Clin Microbiol. 1992 Jul;30(7):1801–1806. [PMC free article] [PubMed]
  • Cravioto A, Tello A, Navarro A, Ruiz J, Villafán H, Uribe F, Eslava C. Association of Escherichia coli HEp-2 adherence patterns with type and duration of diarrhoea. Lancet. 1991 Feb 2;337(8736):262–264. [PubMed]
  • Franke J, Franke S, Schmidt H, Schwarzkopf A, Wieler LH, Baljer G, Beutin L, Karch H. Nucleotide sequence analysis of enteropathogenic Escherichia coli (EPEC) adherence factor probe and development of PCR for rapid detection of EPEC harboring virulence plasmids. J Clin Microbiol. 1994 Oct;32(10):2460–2463. [PMC free article] [PubMed]
  • Frankel G, Riley L, Giron JA, Valmassoi J, Friedmann A, Strockbine N, Falkow S, Schoolnik GK. Detection of Shigella in feces using DNA amplification. J Infect Dis. 1990 Jun;161(6):1252–1256. [PubMed]
  • Karch H, Meyer T. Single primer pair for amplifying segments of distinct Shiga-like-toxin genes by polymerase chain reaction. J Clin Microbiol. 1989 Dec;27(12):2751–2757. [PMC free article] [PubMed]
  • Moseley SL, Echeverria P, Seriwatana J, Tirapat C, Chaicumpa W, Sakuldaipeara T, Falkow S. Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. J Infect Dis. 1982 Jun;145(6):863–869. [PubMed]
  • Nataro JP, Deng Y, Maneval DR, German AL, Martin WC, Levine MM. Aggregative adherence fimbriae I of enteroaggregative Escherichia coli mediate adherence to HEp-2 cells and hemagglutination of human erythrocytes. Infect Immun. 1992 Jun;60(6):2297–2304. [PMC free article] [PubMed]
  • Nataro JP, Kaper JB, Robins-Browne R, Prado V, Vial P, Levine MM. Patterns of adherence of diarrheagenic Escherichia coli to HEp-2 cells. Pediatr Infect Dis J. 1987 Sep;6(9):829–831. [PubMed]
  • Nataro JP, Yikang D, Giron JA, Savarino SJ, Kothary MH, Hall R. Aggregative adherence fimbria I expression in enteroaggregative Escherichia coli requires two unlinked plasmid regions. Infect Immun. 1993 Mar;61(3):1126–1131. [PMC free article] [PubMed]
  • Olive DM. Detection of enterotoxigenic Escherichia coli after polymerase chain reaction amplification with a thermostable DNA polymerase. J Clin Microbiol. 1989 Feb;27(2):261–265. [PMC free article] [PubMed]
  • Savarino SJ, Fasano A, Watson J, Martin BM, Levine MM, Guandalini S, Guerry P. Enteroaggregative Escherichia coli heat-stable enterotoxin 1 represents another subfamily of E. coli heat-stable toxin. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):3093–3097. [PMC free article] [PubMed]
  • Schmidt H, Rüssmann H, Schwarzkopf A, Aleksic S, Heesemann J, Karch H. Prevalence of attaching and effacing Escherichia coli in stool samples from patients and controls. Zentralbl Bakteriol. 1994 Aug;281(2):201–213. [PubMed]
  • Scotland SM, Smith HR, Said B, Willshaw GA, Cheasty T, Rowe B. Identification of enteropathogenic Escherichia coli isolated in Britain as enteroaggregative or as members of a subclass of attaching-and-effacing E. coli not hybridising with the EPEC adherence-factor probe. J Med Microbiol. 1991 Nov;35(5):278–283. [PubMed]
  • Vial PA, Robins-Browne R, Lior H, Prado V, Kaper JB, Nataro JP, Maneval D, Elsayed A, Levine MM. Characterization of enteroadherent-aggregative Escherichia coli, a putative agent of diarrheal disease. J Infect Dis. 1988 Jul;158(1):70–79. [PubMed]

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